Effect of Hepatocyte Growth Factor on the Expression of Matrix Metalloproteinases in the Invasion of Endometrial Cancer Cells in a Three-Dimensional Coculture System

Abstract

Matrix metalloproteinase (MMP)-2 and -9 are secreted and translocated from endometrial stromal cells to cancer cells in a steroid-dependent manner. We investigated the paracrine effect of hepatocyte growth factor (HGF) on the expression of MMPs and tissue inhibitor of metalloproteinases (TIMPs) in stromal and endometrial cancer cells, and correlated the results with cancer cell invasiveness in a three-dimensional (3D) coculture. The 3D coculture of endometrial stromal and cancer cell lines (HEC-1A, HEC-IB, or KLE) was maintained in the presence or absence of HGF. Invasion of the cancer cells was quantified by Boyden’s chamber assay. Under the same conditions, the expression of MMP-2 and -9, membrane type-1 matrix metalloproteinase (MT1-MMP), and TIMP-1 and -2 were examined by zymography and reverse transcription-polymerase chain reaction. A significant increase in the invasiveness of all three cancer cells in the presence of HGF was observed by Boyden’s chamber assay. HGF enhanced the activation of MMP-2 and -9 by zymography of MMPs. HGF strongly induced MMP-9 mRNA expression in stromal cells, but had little effect on MMP-2 mRNA. MT1-MMP mRNA was detected only in KLE and stromal cells, and was also increased by the presence of HGF. TIMP-1 and -2 mRNAs were ubiquitous, with no dependence on HGF.